Organelle motility within mitotic asters of the fungus Nectria haematococca

Abstract

The mitotic asters of the fungus, Nectria haematococca, pull on the spindle pole bodies during anaphase B and help to elongate the central spindle. Because these asters are invisible in vivo, studies of their functions during mitosis have been limited. Invisible asters in other organisms can be studied in vivo because of visible, membranous organelles that are held or transported within them. This is the first report of intra-astral motility of organelles in a fungus, and it lays the foundation for additional studies of aster function in vivo. Using phase-contrast, video-enhanced microscopy, we observed directed motility of mitochondria, small vesicles of various kinds, lipid bodies and, rarely, small vacuoles within the astral region during anaphase B. Both bidirectional motility--toward and away from the spindle pole body--and reversal of direction by an individual organelle were common. Organelles usually did not tend to accumulate either within the aster or near the spindle pole. They were drawn toward the spindle pole body from up to 5.0 microns away. Average velocities were 2.3 to 3.2 microns/s, depending on the organelle and its direction of movement. Transmission electron microscopy revealed apparent cross bridging between astral microtubules and mitochondria, vesicles, endoplasmic reticulum, microbodies, and vacuoles. The antimicrotubule drug, methylbenzimidazole-2-ylcarbamate (MBC), destroyed astral microtubules and virtually eliminated intra-astral motility in vivo, whereas the antiactin drug, cytochalasin E, did not greatly affect the frequency of intra-astral motility episodes. The results suggest a role for astral microtubules in intra-astral motility in this fungus.